RIP-Cre revisited, evidence for impairments of pancreatic beta-cell function

The Cre/loxP recombinase system for performing conditional gene targeting experiments has been very useful in exploring genetic pathways that control both the development and function of pancreatic beta-cells. One particular line of transgenic mice (B6.Cg-Tg(Ins2-cre)25Mgn/J), commonly called RIP-Cre, in which expression of Cre recombinase is controlled by a short fragment of the rat insulin II gene promoter has been used in at least 21 studies on at least 17 genes. In most of these studies inactivation of the gene of interest was associated with either glucose intolerance or frank diabetes. Experimental evidence has been gradually emerging to suggest that RIP-Cre mice alone display glucose intolerance. In this study experiments from three laboratories demonstrate that RIP-Cre mice, in the absence of genes targeted by loxP sites, are glucose intolerant, possibly due to impaired insulin secretion. In addition, we review the use of RIP-Cre mice and discuss possible molecular underpinnings and ramifications of our findings.     

Simultaneous fMRI and Electrophysiology in the Rodent Brain

To examine the neural basis of the blood oxygenation level dependent (BOLD) magnetic resonance imaging (MRI) signal, we have developed a rodent model in which functional MRI data and in vivo intracortical recording can be performed simultaneously. The combination of MRI and electrical recording is technically challenging because the electrodes used for recording distort the MRI images and the MRI acquisition induces noise in the electrical recording. To minimize the mutual interference of the two modalities, glass microelectrodes were used rather than metal and a noise removal algorithm was implemented for the electrophysiology data. In our studies, two microelectrodes were separately implanted in bilateral primary somatosensory cortices (SI) of the rat and fixed in place. One coronal slice covering the electrode tips was selected for functional MRI. Electrode shafts and fixation positions were not included in the image slice to avoid imaging artifacts. The removed scalp was replaced with toothpaste to reduce susceptibility mismatch and prevent Gibbs ringing artifacts in the images. The artifact structure induced in the electrical recordings by the rapidly-switching magnetic fields during image acquisition was characterized by averaging all cycles of scans for each run. The noise structure during imaging was then subtracted from original recordings. The denoised time courses were then used for further analysis in combination with the fMRI data. As an example, the simultaneous acquisition was used to determine the relationship between spontaneous fMRI BOLD signals and band-limited intracortical electrical activity. Simultaneous fMRI and electrophysiological recording in the rodent will provide a platform for many exciting applications in neuroscience in addition to elucidating the relationship between the fMRI BOLD signal and neuronal activity.Download video file.^{(95M, mp4)}     

Harmonised Australian Reference Intervals for Serum PINP and CTX in Adults

Bone turnover markers (BTMs) are classified as either formation or resorption markers. Their concentrations in blood or urine of adults are considered to reflect the rate of bone remodelling and may be of use in the management of patients with bone disease. Major inter-method differences exist for BTMs, and harmonisation of methods is currently being pursued at an international level. Based on published data, this article describes age- and sex-specific Australian consensus reference intervals for adults for serum procollagen type I amino-terminal propeptide (s-PINP) and serum β-isomerised carboxy-terminal cross-linking telopeptide of type I collagen (s-CTX).     

RCPAQAP First Combined Measurement and Reference Interval Survey

Reference intervals are commonly considered to allow for between-laboratory bias. The RCPAQAP Liquid Serum Chemistry Program has collected data on laboratory measurements as well as reference intervals. This allows assessment of the between-laboratory variation in results, reference intervals and the information transmitted by the combination of these factors. For the majority of common chemistry analytes, the between-laboratory variation in reference intervals is greater than the variation in results. Additionally the reference interval variation is generally not related to bias between the results. Use of common reference intervals, either as an average of the current intervals in use, or the intervals proposed by the AACB Harmonisation Group, improved the variation seen in the information produced by different laboratories.     

Landscape spatial patterns in freshwater snail assemblages across Northern Highland catchments

1. Limnologists and landscape ecologists have illustrated how the spatial position of a lake in a landscape influences many of its properties, from the physical to the social. Taking a community ecology perspective, we investigated whether freshwater gastropod assemblages respond to lake landscape position.
2. We determined: (a) whether there is any spatial pattern among lakes in either the species richness or composition of gastropod assemblages; (b) the form of any spatial pattern; and (c) if any explanatory variables (e.g. dispersal corridors and limiting local conditions) show a similar pattern.
3. In three different hydrological catchments, snail species richness increased from isolated highland lakes to stream-connected lowland lakes, probably reflecting increased colonization potential and less limiting local factors for lowland drainage lakes. Catchments appear to differ from one another with regard to relative species abundance, both in terms of macrophyte-associated snail fauna and snails from all habitats aggregated. One or more historical events, such as chance dispersal, may have produced this pattern. Taken together, these results suggest that within-catchment constraints produce repeated gradients in species richness, regardless of what species composition persists in the catchment.     

A geomorphic template for the analysis of lake districts applied to the Northern Highland Lake District, Wisconsin, U.S.A.

1. We tested the degree to which a lake's landscape position constrains the expression of limnological features and imposes a characteristic spatial pattern in a glacial lake district, the Northern Highland Lake District in north‐central Wisconsin. 2. We defined lake order as a metric to analyze the effect of landscape position on limnological features. Lake order, analogous to stream order, is based solely on geographical information and is simple to measure. 3. We examined the strength of the relationship between lake order and a set of 25 variables, which included measures of lake morphometry, water optical properties, major ions, nutrients, biology, and human settlement patterns. 4. Lake order explained a significant fraction of the variance of 21 of the 25 variables tested with ANOVA. The fraction of variance explained varied from 12% (maximum depth) to 56% (calcium concentration). The variables most strongly related to lake order were: measures of lake size and shape, concentrations of major ions (except sulfate) and silica, biological variables (chlorophyll concentration, crayfish abundance, and fish species richness), and human‐use variables (density of cottages and resorts). Lake depth, water optical properties, and nutrient concentrations (other than silica) were poorly associated with lake order. 5. Potential explanations for a relationship with lake order differed among variables. In some cases, we could hypothesize a direct link. For example, major ion concentration is a function of groundwater input, which is directly related to lake order. We see these as a direct influence of the geomorphic template left by the retreat of the glacier that led to the formation of this lake district. 6. In other cases, a set of indirect links was hypothesized. For example, the effect of lake order on lake size, water chemistry, and lake connectivity may ultimately explain the relation between lake order and fish species richness. We interpret these relationships as the result of constraints imposed by the geomorphic template on lake development over the last 12 000 years. 7. By identifying relationships between lake characteristics and a measure of landscape position, and by identifying geomorphologic constraints on lake features and lake evolution, our analysis explains an important aspect of the spatial organization of a lake district.     

Trichloroethene Reductive Dehalogenase from Dehalococcoides ethenogenes: Sequence of tceA and Substrate Range Characterization

The anaerobic bacterium Dehalococcoides ethenogenes is the only known organism that can completely dechlorinate tetrachloroethene or trichloroethene (TCE) to ethene via dehalorespiration. One of two corrinoid-containing enzymes responsible for this pathway, TCE reductive dehalogenase (TCE-RDase) catalyzes the dechlorination of TCE to ethene. TCE-RDase dehalogenated 1,2-dichloroethane and 1,2-dibromoethane to ethene at rates of 7.5 and 30 μmol/min/mg, respectively, similar to the rates for TCE, cis-dichloroethene (DCE), and 1,1-DCE. A variety of other haloalkanes and haloalkenes containing three to five carbon atoms were dehalogenated at lower rates. The gene encoding TCE-RDase, tceA, was cloned and sequenced via an inverse PCR approach. Sequence comparisons of tceA to proteins in the public databases revealed weak sequence similarity confined to the C-terminal region, which contains the eight-iron ferredoxin cluster binding motif, (CXXCXXCXXXCP)₂. Direct N-terminal sequencing of the mature enzyme indicated that the first 42 amino acids constitute a signal sequence containing the twin-arginine motif, RRXFXK, associated with the Sec-independent membrane translocation system. This information coupled with membrane localization studies indicated that TCE-RDase is located on the exterior of the cytoplasmic membrane. Like the case for the two other RDases that have been cloned and sequenced, a small open reading frame, tceB, is proposed to be involved with membrane association of TCE-RDase and is predicted to be cotranscribed with tceA.